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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Human genetics 〈Berlin〉 28 (1975), S. 295-302 
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung In Kulturen menschlicher Leukocyten mit TC medium 199 wurden die DNS-Synthese und die Mitoseindices mit Hilfe von 3H-Thymidin-Autoradiographie und Zellzählung bestimmt. Die DNS-Synthese beginnt bei etwa 28 Std. Die Frequenzen markierter Zellen nehmen langsam zu und erreichen Maximalwerte von etwa 24%. Die ersten Mitosen erscheinen bei etwa 38 Std. Bis 49 Std werden nur wenige Mitosen beobachtet, danach nehmen die Mitoseindices zu und erreichen Werte bis etwa 11‰. Kultivierung mit BUdR und Fluorescenzfärbung mit Hoechst 33258-Farbstoff ergab, daß nach 72 Std in den Kulturen erste, zweite und dritte Mitosen nebeneinander vorkommen. Bestrahlung von Vollblut und anschließende Kultivierung für 72 Std führt zu Mitosen mit dizentrischen Chromosomen und Ringchromosomen mit und ohne Fragmente, zu Interphasen mit Mikrokernen, zu vorzeitig kondensiertem Chromatin und zu Endoreduplikationen und tetraploiden Mitosen. Dieser Befund zeigt ebenfalls, daß nach 72 Std erste und weitere Mitosen gemeinsam in den Kulturen vorkommen.
    Notes: Summary In human leukocyte cultures set up with TC medium 199, DNA synthesis and mitotic indices were analysed by means of 3H-thymidine autoradiography and cell counting. DNA synthesis starts at around 28 hrs. The frequencies of labelled cells rise slowly and reach a maximum of around 24%. The first mitoses appear at around 38 hrs but up to 49 hrs only very few mitoses can be seen. After that time the mitotic indices rise and reach values of up to 11‰. Cultivation in the presence of BUdR for 72 hrs and staining with Hoechst 33258 stain revealed that first, second and third mitoses occur together in the cultures at this time. Irradiation of whole blood and cultivation for 72 hrs leads to mitoses containing dicentric and ring chromosomes with an without fragments, to interphases with micronuclei, to premature chromosome condensations (PCC) and to polyploid mitoses indicating that at this time first and further mitoses are present.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Chromosomes were isolated from Chinese hamster x human hybrid cell lines containing four and nine human chromosomes. Human genomic DNA was biotinylated by nick translation and used to label the human chromosomes by in situ hybridization in suspension. Streptavidin was covalently coupled to the surface of magnetic beads and these were incubated with the hybridized chromosomes. The human chromosomes were bound to the magnetic beads through the strong biotin-streptavidin complex and then rapidly separated from nonlabeled Chinese hamster chromosomes by a simple permanent magnet. The hybridization was visualized by additional binding of avidin-FITC (fluorescein) to the unoccupied biotinylated human DNA bound to the human chromosomes. After magnetic separation, up to 98% of the individual chromosomes attached to magnetic beads were classified as human chromosomes by fluorescence microscopy.
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  • 3
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A procedure was developed to provide differential fluorescent staining of metaphase chromosomes in suspension following nucleic acid hybridization. For this purpose metaphase chromosomes were isolated from a Chinese hamster x human hybrid cell line. After hybridization with biotinylated human genomic DNA, the human chromosomes were visualized by indirect immunofluorescence using antibodies against biotin and fluoresceine-isothiocyanate-(FITC)-labeled second antibodies. This resulted in green fluorescent human chromosomes. In contrast, Chinese hamster chromosomes revealed red fluorescent staining only when counterstained with propidium iodide. Notably, interspecies chromosomal rearrangements could be easily detected. After hybridization and fluorescent staining, chromosomes still showed a well-preserved morphology under the light microscope. We suggest that this procedure may have a useful application in flow cytometry and sorting.
    Type of Medium: Electronic Resource
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